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Methods Cell Biol. 2019;154:1-14. doi: 10.1016/bs.mcb.2019.03.010. Epub 2019 Apr 24.

Single glomerular proteomics: A novel tool for translational glomerular cell biology.

Author information

1
The Scripps Research Institute, Center for Mass Spectrometry and Metabolomics, San Diego, CA, United States; Department of Internal Medicine II and Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany. Electronic address: markus.rinschen@uk-koeln.de.

Abstract

The glomerulus harbors the renal filtration barrier and needs to be precisely maintained. In this chapter, the concept of single glomerular proteomics is described. Single glomerular proteomics has recently been enabled by advances in glomerular isolation, ultrasensitive peptide sample preparation and mass spectrometry based technology and acquisition strategies. It generates protein content information on a single glomerulus that can be overlaid with morphological and other multi-layered omics analyses. The novel method consists of four essential steps: preparation of single glomeruli-by microdissection, glomerular preparation, or laser microdissection-followed by proteomic sample preparation, mass spectrometry analysis and bioinformatics analysis. It enables for the first time the generation of sub-biopsy level proteomics data. In perspective, comprehensive data from individual glomeruli could be used in order to pinpoint novel druggable targets in animal models of kidney disease or in patients with proteinuria and glomerular disease.

KEYWORDS:

Diabetic nephropathy; Endothelium; Focal segmental glomerular sclerosis; Hypertensive kidney disease; Mesangial cells; Podocyte; SP3; Single nephron; Ultrasensitive proteomics

PMID:
31493812
DOI:
10.1016/bs.mcb.2019.03.010

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